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Protein Secretion [electronic resource] : Methods and Protocols / edited by Anastassios Economou.

Edition:
1.
Publication:
Totowa, NJ : Humana Press, 2010.
Series:
Methods in Molecular Biology, Methods and Protocols, 1064-3745 ; 619
Springer Protocols (Springer-12345)
Methods in Molecular Biology, Methods and Protocols, 1064-3745 ; 619
Format/Description:
Book
1 online resource (XV, 426 pages) : 74 illustrations, 2 illustrations in color.
Subjects:
Life sciences.
Biochemistry.
Proteomics.
Enzymology.
Local subjects:
Life Sciences.
Enzymology.
Biochemistry, general.
Proteomics.
System Details:
text file PDF
Summary:
Over the past thirty years, many elegant genetic and biochemical approaches have been combined in order to advance the study of protein secretion and the necessary navigation through cell membranes, yet, despite this progress, less than two hundred membrane protein structures are known, nowhere near the complete inventory that the discovered protein export systems suggest. In Protein Secretion: Methods and Protocols, leading experts in the field provide robust, well-established protocols to elucidate the multiplicity of tools that have been developed to study protein sorting, membrane targeting, transmembrane crossing, and secretion across multiple membranes. With examples involving both prokaryotic and eukaryotic organisms, the volume covers subjects ranging from bioinformatics and proteomics to fundamental enzymology and genetics to cell biology, structural analyses, and biophysics. Written in the highly successful Methods in Molecular Biology™ series format, chapters contain introductions to their respective topics, lists of the key materials and reagents, step-by-step, readily reproducible protocols, and detailed notes on troubleshooting and avoiding known pitfalls. Comprehensive and dependable, Protein Secretion: Methods and Protocols focuses on well-characterized paradigms so that scientists studying a vast array of subjects from biochemistry and genetics to biotechnology and biopharmaceuticals can benefit and expand upon their vital research.
Contents:
The Extraordinary Diversity of Bacterial Protein Secretion Mechanisms
In Vitro and In Vivo Approaches to Studying the Bacterial Signal Peptide Processing
Membrane Insertion of Small Proteins
Membrane Protein Insertion in E. coli
Study of Polytopic Membrane Protein Topological Organization as a Function of Membrane Lipid Composition
In Vivo Analysis of Protein Translocation to the Escherichia coli Periplasm
Sorting of Bacterial Lipoproteins to the Outer Membrane by the Lol System
Purification and Functional Reconstitution of the Bacterial Protein Translocation Pore, the SecYEG Complex
Reconstitution of the SecY Translocon in Nanodiscs
In Vitro Assays to Analyze Translocation of the Model Secretory Preprotein Alkaline Phosphatase
Characterization of Interactions Between Proteins Using Site-Directed Spin Labeling and Electron Paramagnetic Resonance Spectroscopy
Analysis of Tat Targeting Function and Twin-Arginine Signal Peptide Activity in Escherichia coli
Site-Specific Cross-Linking of In Vitro Synthesized E. coli Preproteins for Investigating Transmembrane Translocation Pathways
Tracking the Secretion of Fluorescently Labeled Type III Effectors from Single Bacteria in Real Time
Comparative Analysis of Cytoplasmic Membrane Proteomes of Escherichia coli Using 2D Blue Native/SDS-PAGE
Using Hidden Markov Models to Discover New Protein Transport Machines
Bioinformatics Predictions of Localization and Targeting
The Chloroplast Protein Import Machinery: A Review
Measurement of the Energetics of Protein Transport Across the Chloroplast Thylakoid Membrane
In Vitro Dissection of Protein Translocation into the Mammalian Endoplasmic Reticulum
In Vitro Reconstitution of the Selection, Ubiquitination, and Membrane Extraction of a Polytopic ERAD Substrate
Studying the ArfGAP-Dependent Conformational Changes in SNAREs
Studying Endoplasmic Reticulum Function In Vitro Using siRNA
High-Quality Immunofluorescence of Cultured Cells
Trapping Oxidative Folding Intermediates During Translocation to the Intermembrane Space of Mitochondria: In Vivo and In Vitro Studies
Native Techniques for Analysis of Mitochondrial Protein Import.
Contributor:
Economou, Anastassios. editor.
SpringerLink (Online service)
Contained In:
Springer eBooks
Other format:
Printed edition:
ISBN:
9781603274128
Publisher Number:
10.1007/978-1-60327-412-8 doi
Access Restriction:
Restricted for use by site license.
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